Identification and characterization of a selenoprotein, thioredoxin reductase, in a unicellular marine haptophyte alga, Emiliania huxleyi.
We found six selenoproteins (EhSEP1-6) in the coccolithophorid Emiliania huxleyi (Haptophyceae) using the (75)Se radiotracer technique. Previously, the most abundant selenoprotein, EhSEP2, was identified as a novel selenoprotein, a protein disulfide isomerase-like protein (Obata, T., and Shiraiwa, Y. (2005) J. Biol. Chem. 280, 18462-18468). The present study focused on the second abundant selenoprotein, EhSEP1, in the same cells and analyzed its molecular properties and regulation of gene expression by selenium. The cDNA sequence of EhSEP1 consists of 1950 base pairs encoding a putative product of 495 amino acids with a calculated molecular mass of 52.2 kDa. The nucleotide and amino acid sequences of EhSEP1 showed strong similarities to those of the enzyme thioredoxin reductase (TR) 1 in the public databases. The EhSEP1 protein contains redox-active cysteine residues in the putative FAD binding domain of the pyridine nucleotide-disulfide oxidoreductase class-1 domain, a dimerization domain, and a C-terminal Gly-Cys-Sec (selenocysteine)-Gly sequence that is known to function as an additional redox center. In the 3'-untranslated region of EhSEP1 cDNA, we found a selenocysteine insertion sequence (SECIS) that is similar to the SECIS found previously in animals. The expression of EhSEP1 showed almost the same pattern under both selenium-sufficient and selenium-deficient conditions. Conversely, TR activity gradually increased 4-fold within ca. 70 h when cells were transferred to the medium containing 10 nm selenite. These data show that selenium is essential for the induction of TR activity at the translational level but not at the transcriptional level in this alga.
Araie
Hiroya
H
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572, Japan.
Suzuki
Iwane
I
Shiraiwa
Yoshihiro
Y
D020418:Algal Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D056890:Eukaryota / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D051140:Selenoproteins / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology
D020418:Algal Proteins; D018076:DNA, Complementary; D051140:Selenoproteins; D013880:Thioredoxin-Disulfide Reductase